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PCR Free Detection of Hepatitis B Virus DNA Using a Nanostructured Impedance Biosensor

[+] Author Affiliations
Chun-Cheng Chen, Gou-Jen Wang

National Chung-Hsing University, Taichung, Taiwan

Paper No. DETC2014-34866, pp. V004T09A004; 6 pages
doi:10.1115/DETC2014-34866
From:
  • ASME 2014 International Design Engineering Technical Conferences and Computers and Information in Engineering Conference
  • Volume 4: 19th Design for Manufacturing and the Life Cycle Conference; 8th International Conference on Micro- and Nanosystems
  • Buffalo, New York, USA, August 17–20, 2014
  • Conference Sponsors: Design Engineering Division, Computers and Information in Engineering Division
  • ISBN: 978-0-7918-4635-3
  • Copyright © 2014 by ASME

abstract

In this study, a PCR free technique for effective detection of hepatitis B virus (HBV) DNA obtained directly from clinical samples was presented. The honeycomb-like barrier layer of an anodic aluminum oxide (AAO) film having a uniform nanohemisphere array was used as the substrate of the sensing electrode. A gold thin film about 30 nm thick was radio-frequency (RF) magnetron sputtered onto the AAO barrier-layer surface as the electrode for the successive deposition of gold nanoparticles (GNPs) on the hemisphere surface. A specially designed single-strand 96-mer gene fragment of the target genomic DNA of HBV based on the genome sequences of HBV from the National Center for Biotechnology Information (NCBI) was immobilized on the nanostructured electrode as the capture probe. Complementary target HBV DNA (3020–3320 mer) obtained from clinical samples were further hybridized to the sensing probes. Detection results through electrochemical impedance spectroscopy (EIS) illustrate that two dynamic linear ranges, 0–103 and 103–105 copies/mL, having R2 values of 0.973 and 0.998, respectively, could be obtained. A detection limit of 186 copies/mL could be achieved. The proposed simple and high performance HBV DNA detection technique in this study is highly feasible for future clinical applications.

Copyright © 2014 by ASME
Topics: Biosensors , DNA

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